Skip to main content

Advertisement

SpringerLink
Log in

RP-HPLC Method for Simultaneous Determination of Sofosbuvir and Ledipasvir in Tablet Dosage Form and Its Application to In Vitro Dissolution Studies

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

A reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sofosbuvir and ledipasvir in tablet dosage form. The analysis was performed on Luna analytical column 250 × 4.6 mm, 5 µm, octyl silica packing (Si–[CH2]7–CH3) C8, using ammonium acetate buffer solution pH 7.0 and acetonitrile 35:65 % v/v as mobile phase at flow rate of 0.7 mL min−1 for isocratic elution. Detection of sofosbuvir and ledipasvir was performed on a UV detector at 245 nm. The retention times of sofosbuvir and ledipasvir were 4.468 ± 0.013 min and 8.242 ± 0.012 min, respectively, and the total run time was 20 min. The method was validated according to the requirements of the United States Pharmacopeia (category I). The overall recovery of both analytes was 100 ± 1 %; the relative standard deviation for precision and intraday precision was less than 2.0 %. The method was linear with correlation coefficient (r) >0.9999, limits of detection 0.485 and 0.175 µg mL−1, and limits of quantification was 1.619 and 0.586 µg mL−1 for sofosbuvir and ledipasvir, respectively. The method was successfully applied to the assay and in vitro dissolution studies of sofosbuvir and ledipasvir in tablet dosage form.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Lavanchy D (2011) Evolving epidemiology of hepatitis C virus. Clin Microbiol Infect 17:107–115

    Article  CAS  Google Scholar 

  2. Shepard CW, Finelli L, Alter MJ (2005) Global epidemiology of hepatitis C virus infection. Lancet Infect Dis 5:558–567

    Article  Google Scholar 

  3. Jacobson IM, Davis GL, EI-Serag H, Negro F, Trepo C (2010) Prevalence and challenges of liver diseases in patients with chronic hepatitis C virus infection. Clin Gastroenterol Hepatol 8:924–933

    Article  Google Scholar 

  4. Poynard T, Afdhal NH (2010) Perspectives on fibrosis progression in hepatitis C: ana la carte approach to risk factors and staging of fibrosis. Antivir Ther 15:281–291

    Article  CAS  Google Scholar 

  5. Naggie S, Patel K, McHutchison J (2010) Hepatitis C virus directly acting antivirals: current development with NS3/4A HCV serine protease inhibitors. J Antimicrob Chemother 65:2063–2269

    Article  CAS  Google Scholar 

  6. Magiorkinis G, Sypsa V, Magiorkinis E, Paraskevis D, Katsoulidou A, Belshaw R, Fraser C, Pybus OG, Hatzakis A (2013) Integrating phylodynamics and epidemiology to estimate transmission diversity in viral epidemics. PLoS Comput Biol 9:e1002876

    Article  CAS  Google Scholar 

  7. Pearlman BL, Traub N (2011) Sustained virologic response to antiviral therapy for chronic hepatitis C virus infection: a cure and so much more. Clin Infect Dis 52:889–900

    Article  Google Scholar 

  8. Ghany MG, Nelson DR, Strader DB, Thomas DL, Seeff LB (2011) An update on treatment of genotype 1 chronic hepatitis C virus infection: 2011 practice guideline by the American Association for the Study of Liver Diseases. Hepatology 54:1433–1444

    Article  Google Scholar 

  9. Harrison SA (2012) Management of anemia in patients receiving protease inhibitors. Gastroenterol Hepatol 8:254–256

    Google Scholar 

  10. Jacobson IM, McHutchison JG, Dusheiko G, Di Bisceglie AM, Reddy KR, Bzowej NH, Marcellin P, Muir AJ, Ferenci P, Flisiak R, George J, Rizzetto M, Shouval D, Sola R, Terg RA, Yoshida EM, Adda N, Bengtsson L, Sankoh AJ, Kieffer TL, George S, Kauffman RS, Zeuzem S (2011) Telaprevir for previously untreated chronic hepatitis C virus infection. N Engl J Med 364:2405–2416

    Article  CAS  Google Scholar 

  11. Poordad F, McCone J Jr, Bacon BR, Bruno S, Manns MP, Sulkowski MS, Jacobson IM, Reddy KR, Goodman ZD, Boparai N, DiNubile MJ, Sniukiene V, Brass CA, Albrecht JK, Bronowicki JP (2011) Boceprevir for untreated chronic HCV genotype 1 infection. N Engl J Med 364:1195–1206

    Article  CAS  Google Scholar 

  12. Cholongitas E, Papatheodoridis GV (2014) Sofosbuvir: a novel oral agent for chronic hepatitis C. Ann Gastroenterol 27:331–337

    Google Scholar 

  13. Keating GM, Vaidya A (2014) Sofosbuvir: first global approval. Drugs 74:273–282

    Article  CAS  Google Scholar 

  14. Rose L, Bias TE, Mathias CB, Trooskin SB, Fong JJ (2014) Sofosbuvir: a nucleotide NS5B inhibitor for the treatment of chronic hepatitis C infection. Ann Pharmacother 48:1019–1029

    Article  Google Scholar 

  15. Link JO, Taylor JG, Xu L, Mitchell M, Guo H, Liu H, Kato D, Kirschberg T, Sun J, Squires N, Parrish J, Keller T, Yang ZY, Yang C, Matles M, Wang Y, Wang K, Cheng G, Tian Y, Mogalian E, Mondou E, Cornpropst M, Perry J, Desai MC (2014) Discovery of ledipasvir (GS-5885): a potent, once-daily oral NS5A inhibitor for the treatment of hepatitis C virus infection. J Med Chem 57:2033–2046

    Article  CAS  Google Scholar 

  16. Everson G, Lawitz E, Thompson, Sulkowski M, Zhu Y, Brainard, D, Mendelson L, McHutchison J, Pang PS, Yang JC, Marcellin P, Afdhal N (2012) The NS5A inhibitor GS-5885 is safe and well tolerated in over 1000 patients treated in phase 2 studies: 63rd annual meeting of the American Association for the study of liver diseases, Boston. http://www.natap.org/2012/AASLD/AASLD_04.htm. Accessed 18 Aug 2015

  17. Afdhal N, Zeuzem S, Kwo P, Chojkier M, Gitlin N, Puoti M, Romero-Gomez M, Zarski JP, Agarwal K, Buggisch P, Foster GR, Bräu N, Buti M, Jacobson IM, Subramanian GM, Ding X, Mo H, Yang JC, Pang PS, Symonds WT, McHutchison JG, Muir AJ, Mangia A, Marcellin P, ION-1 Investigators (2014) Ledipasvir and sofosbuvir for untreated HCV genotype 1 infection. N Engl J Med 370:1889–1898

    Article  Google Scholar 

  18. Kowdley KV, Gordon SC, Reddy KR, Rossaro L, Bernstein DE, Lawitz E, Shiffman ML, Schiff E, Ghalib R, Ryan M, Rustgi V, Chojkier M, Herring R, Di Bisceglie AM, Pockros PJ, Subramanian GM, An D, Svarovskaia E, Hyland RH, Pang PS, Symonds WT, McHutchison JG, Muir AJ, Pound D, Fried MW, ION-3 Investigators (2014) Ledipasvir and sofosbuvir for 8 or 12 weeks for chronic HCV without cirrhosis. N Engl J Med 370:1879–1888

    Article  Google Scholar 

  19. Amidon GL, Lennernäs H, Shah VP, Crison JR (1995) A theoretical basis for a biopharmaceutic drug classification: the correlation of in vitro drug product dissolution and in vivo bioavailability. Pharm Res 12:413–420

    Article  CAS  Google Scholar 

  20. Assessment report EMA/CHMP/702742/2014 (2014) Committee for Medicinal Products for Human Use (CHMP), London. http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Public_assessment_report/human/003850/WC500177996.pdf. Accessed 18 Aug 2015

  21. Khan KA (1975) The concept of dissolution efficiency. J Pharm Pharmacol 28:48–49

    Article  Google Scholar 

  22. Devane J, Butler J (1997) The impact of in vitro–in vivo relationships on product development. Pharm Technol 21:146–159

    Google Scholar 

  23. Emami J (2006) In vitro–in vivo correlation: from theory to applications. J Pharm Pharm Sci 9:169–189

    CAS  Google Scholar 

  24. Azarmi S, Roa W, Löbenberg R (2007) Current perspectives in dissolution testing of conventional and novel dosage forms. Int J Pharm 328:12–21

    Article  CAS  Google Scholar 

  25. Rezk MR, Basalious EB, Karim IA (2015) Development of a sensitive UPLC–ESI–MS/MS method for quantification of sofosbuvir and its metabolite GS-331007 in human plasma: application to a bioequivalence study. J Pharm Biomed Anal 114:97–104

    Article  CAS  Google Scholar 

  26. Qu L, Wang W, Zeng D, Lu Y, Yin Z (2015) Quantitative performance of online SPE-LC coupled to Q-Exactive for the analysis of sofosbuvir in human plasma. RSC Adv 5:98269–98277

    Article  CAS  Google Scholar 

  27. Shi X, Zhu D, Lou J, Zhu B, Hu AR, Gan D (2015) Evaluation of a rapid method for the simultaneous quantification of ribavirin, sofosbuvir and its metabolite in rat plasma by UPLC–MS/MS. J Chromatogr B 1002:353–357

    Article  CAS  Google Scholar 

  28. Rower JE, Jimmerson LC, Chen X, Zheng J-H, Hodara A, Bushman LR, Anderson PL, Kiser JJ (2015) Validation and application of a liquid chromatography-tandem mass spectrometry method to determine the concentrations of sofosbuvir anabolites in cells. Antimicrob Agents Chemother 59:7671–7679

    Article  CAS  Google Scholar 

  29. US FDA, Center for Drug Evaluation and Research (CDER), Generic drugs division of bioequivalence: FDA-recommended dissolution methods. http://www.accessdata.fda.gov/scripts/cder/dissolution/dsp_getallData.cfm. Accessed 21 Nov 2015

  30. US FDA, Center for Drug Evaluation and Research (CDER) (1997) Dissolution testing of immediate release solid oral dosage forms. http://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm070237.pdf. Accessed 3 Mar 2007

  31. USP (2014) The United States Pharmacopeia, 37th revision. United States Pharmacopeial Convention, Rockville, pp 1157–1162

  32. ICH (2005) ICH Harmonised Tripartite Guideline Q2(R1): Validation of analytical procedures. ICH, Geneva. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf. Accessed 3 Mar 2007

Download references

Acknowledgments

The authors are grateful to Genome Pharmaceuticals for their support in this work.

Author information

Authors and Affiliations

  1. Institute of Chemical Sciences, University of Peshawar, Peshawar, 25120, Khyber Pakhtunkhwa, Pakistan

    Bakht Zaman & Waseem Hassan

  2. Department of Pharmacy, University of Peshawar, Peshawar, 25120, Khyber Pakhtunkhwa, Pakistan

    Faisal Siddique

Authors
  1. Bakht Zaman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Faisal Siddique
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Waseem Hassan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Waseem Hassan.

Ethics declarations

Conflict of Interest

The authors declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zaman, B., Siddique, F. & Hassan, W. RP-HPLC Method for Simultaneous Determination of Sofosbuvir and Ledipasvir in Tablet Dosage Form and Its Application to In Vitro Dissolution Studies. Chromatographia 79, 1605–1613 (2016). https://doi.org/10.1007/s10337-016-3179-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-016-3179-9

Keywords

Search

Navigation